Amplicon size for BSP - (Sep/01/2008 )
Hi!!
Does anyone know the optimal amplicon size when you perform a bisulphite sequencing??
Thanx!!
-yarince-
Depends on the quality and fragmentation of your starting DNA, as it is further degraded after bisulphite.
Having said this, 300-400bp is usually fine, the smaller the assay however the better it is!
Nick
-methylnick-
QUOTE (methylnick @ Sep 1 2008, 11:39 PM)
Depends on the quality and fragmentation of your starting DNA, as it is further degraded after bisulphite.
Having said this, 300-400bp is usually fine, the smaller the assay however the better it is!
Nick
Having said this, 300-400bp is usually fine, the smaller the assay however the better it is!
Nick
I've designed primers to generate amplicons of 250-300 pb so I hope it works ok...
Thank you!
-yarince-
I've just got some good sequencing results from a 600bp amplicon.
Remember that you won't get any sequencing results for the first 50 or so bases after the sequencing primer, so give it a little more room if your CpG's are near to the edges.
Good luck!
-Davo-